British Columbia beekeeping revenues and costs: survey data and profit modeling.

British Columbia beekeepers, like many beekeepers around the world, are currently facing declines in honey bee health and high overwinter colony losses. To better understand the economics and the cycle of yearly colony loss and replacement of this critical agricultural industry, we collected and analyzed survey data on beekeeping costs and returns. Forty British Columbia beekeepers provided details about revenue sources, variable costs, capital costs, and investments. Ten surveyed beekeepers managed between 1 and 9 colonies, 10 managed between 10 and 39 colonies, 9 managed between 40 and 100 colonies, 5 managed between 101 and 299 colonies, 3 managed between 300 and 699 colonies, and 3 managed 700 colonies or more. The data was used to calculate beekeeping profit and to parameterize a model that explores the economic impact of colony loss rates and replacement strategies. Survey results show that when the data is aggregated, revenues exceed costs for beekeeping operations in British Columbia with a per colony profit of $56.92 or $0.87 per pound of honey produced. Surveyed operations with fewer than 100 colonies have negative profits, while operations with 100-299 colonies have positive profits. Surveyed operations in the Cariboo, North Coast, and Okanagan regions have the highest profits while surveyed operations in the Peace region have the lowest profits. Profit modeling shows that replacing losses with packages generates lower profit than replacing losses with split colonies. Our modeling shows that operations that diversify their revenue to include bee sales and commercial pollination accrue higher profits and can withstand higher winter loss rates.

Evidence of presence and replication of honey bee viruses among wild bee pollinators in subtropical environments.

We determined the presence of six viruses in different bee species collected in subtropical environments. Deformed wing virus (DWV) and black queen cell virus (BQCV) were detected in >90% of honey bee samples and in 50-100% of four stingless bee, two bumble bee and one solitary bee species. Additionally, minus DWV and BQCV RNA strands were detected, indicating that the viruses replicate in several hosts. This is the first report of honey bee viruses replicating in six wild bee species in the tropics. If pathogenic to them, viral infections could result in negative impacts in agricultural and unmanaged ecosystems.

Viral Quantification in Bee Samples Using Synthetic DNA Sequences with Real-Time PCR (qPCR).

Pathogen spillover between honey bees and wild pollinators is a relatively new and exciting field of study. It is known that some viral diseases are a major threat to honey bee health and, thus, the diagnosis and quantification of honey bee viruses in wild pollinators have gained attention. Pathogen spillover from honey bees to wild bees and the consequences of viral replication to their health still need to be investigated. However, finding positive samples to produce standard curves and include positive controls in real-time PCR (qPCR) assays is challenging. Here we describe the use of synthetic DNA sequences of two variants of deformed wing virus (DWV-A and DWV-B), black queen cell virus (BQCV), sacbrood virus (SBV), chronic bee paralysis virus (CBPV), Kashmir bee virus (KBV), acute bee paralysis virus (ABPV), and Israeli acute paralysis virus (IAPV), to construct standard curves for viral quantification, and for their use as positive controls in qPCR assays.

Breeding honey bees (Apis mellifera L.) for low and high Varroa destructor population growth: Gene expression of bees performing grooming behavior.

Introduction: Social organisms, including honey bees (Apis mellifera L.), have defense mechanisms to control the multiplication and transmission of parasites and pathogens within their colonies. Self-grooming, a mechanism of behavioral immunity, seems to contribute to restrain the population growth of the ectoparasitic mite Varroa destructor in honey bee colonies. Because V. destructor is the most damaging parasite of honey bees, breeding them for resistance against the mite is a high priority of the beekeeping industry. Methods: A bidirectional breeding program to select honey bee colonies with low and high V. destructor population growth (LVG and HVG, respectively) was conducted. Having high and low lines of bees allowed the study of genetic mechanisms underlying self-grooming behavior between the extreme genotypes. Worker bees were classified into two categories: 'light groomers' and 'intense groomers'. The brains of bees from the different categories (LVG-intense, LVG-light, HVG-intense, and HVG-light) were used for gene expression and viral quantification analyses. Differentially expressed genes (DEGs) associated with the LVG and HVG lines were identified. Results: Four odorant-binding proteins and a gustatory receptor were identified as differentially expressed genes. A functional enrichment analysis showed 19 enriched pathways from a list of 219 down-regulated DEGs in HVG bees, including the Kyoto Encyclopedia of Genes and Genomes (KEGG) term of oxidative phosphorylation. Additionally, bees from the LVG line showed lower levels of Apis rhabdovirus 1 and 2, Varroa destructor virus -1 (VDV-1/DWV-B), and Deformed wing virus-A (DWV-A) compared to bees of the HVG line. The difference in expression of odorant-binding protein genes and a gustatory receptor between bee lines suggests a possible link between them and the perception of irritants to trigger rapid self-grooming instances that require the activation of energy metabolic pathways. Discussion: These results provide new insights on the molecular mechanisms involved in honey bee grooming behavior. Differences in viral levels in the brains of LVG and HVG bees showed the importance of investigating the pathogenicity and potential impacts of neurotropic viruses on behavioral immunity. The results of this study advance the understanding of a trait used for selective breeding, self-grooming, and the potential of using genomic assisted selection to improve breeding programs.

The Fungus Nosema ceranae and a Sublethal Dose of the Neonicotinoid Insecticide Thiamethoxam Differentially Affected the Health and Immunity of Africanized Honey Bees.

Honey bees (Apis mellifera L.) are affected by different biotic and abiotic stressors, such as the fungus Nosema ceranae and neonicotinoid insecticides, that negatively impact their health. However, most studies so far conducted have focused on the effect of these stressors separately and in European honey bees. Therefore, this study was conducted to analyze the impact of both stressors, singly and in combination, on honey bees of African descent that have demonstrated resistance to parasites and pesticides. Africanized honey bees (AHBs, Apis mellifera scutellata Lepeletier) were inoculated with N. ceranae (1 × 105 spores/bee) and/or chronically exposed for 18 days to a sublethal dose of thiamethoxam (0.025 ng/bee) to evaluate their single and combined effects on food consumption, survivorship, N. ceranae infection, and immunity at the cellular and humoral levels. No significant effects by any of the stressors were found for food consumption. However, thiamethoxam was the main stressor associated to a significant decrease in AHB survivorship, whereas N. ceranae was the main stressor affecting their humoral immune response by upregulating the expression of the gene AmHym-1. Additionally, both stressors, separately and combined, significantly decreased the concentration of haemocytes in the haemolymph of the bees. These findings indicate that N. ceranae and thiamethoxam differentially affect the lifespan and immunity of AHBs and do not seem to have synergistic effects when AHBs are simultaneously exposed to both stressors.

First insights into the honey bee (Apis mellifera) brain lipidome and its neonicotinoid-induced alterations associated with reduced self-grooming behavior.

Introduction: Honey bees (Apis mellifera) play key roles in food production performing complex behaviors, like self-grooming to remove parasites. However, the lipids of their central nervous system have not been examined, even though they likely play a crucial role in the performance of cognitive process to perform intricate behaviors. Lipidomics has greatly advanced our understanding of neuropathologies in mammals and could provide the same for honey bees. Objectives: The objectives of this study were to characterize the brain lipidome of adult honey bees and to assess the effect of clothianidin (a neurotoxic insecticide) on the brain lipid composition, gene expression, and performance of self-grooming behavior under controlled conditions (cage experiments). Methods: After seven days of exposure to oral sublethal doses of clothianidin, the bees were assessed for self-grooming behavior; their brains were dissected to analyze the lipidome using an untargeted lipidomics approach and to perform a high throughput RNAseq analysis. Results: Compared to all other organisms, healthy bee brain lipidomes contain unusually high levels of alkyl-ether linked (plasmanyl) phospholipids (51.42%) and low levels of plasmalogens (plasmenyl phospholipids; 3.46%). This could make it more susceptible to the effects of toxins in the environment. A positive correlation between CL 18:3/18:1/14:0/22:6, TG 6:0/11:2/18:1, LPE 18:0e and intense self-grooming was found. Sublethal doses of a neonicotinoid altered PC 20:3e/15:0, PC 16:0/18:3, PA 18:0/24:1, and TG 18:1/18:1/18/1 levels, and affected gene expression linked to GPI-anchor biosynthesis pathway and energy metabolism that may be partially responsible for the altered lipid composition. Conclusion: This study showed that lipidomics can reveal honey bee neuropathologies associated with reduced grooming behavior due to sublethal neonicotinoid exposure. The ease of use, unusual brain lipidome as well as characterized behaviors that are affected by the environment make honey bees a promising model organism for studying the neurolipidome and associations with neurobehavioral disorders.

Genotype, but Not Climate, Affects the Resistance of Honey Bees (Apis mellifera) to Viral Infections and to the Mite Varroa destructor.

This study was conducted to analyze the effect of genotype and climate on the resistance of honey bee (Apis mellifera) colonies to parasitic and viral diseases. The prevalence and intensity of parasitism by Varroa destructor, or infection by Nosema spp., and four honey bee viruses were determined in 365 colonies of predominantly European or African ancestry (descendants of A. m. scutellata) in subtropical and temperate regions of Mexico. Varroa destructor was the most prevalent parasite (95%), whilst N. ceranae was the least prevalent parasite (15%). Deformed wing virus (DWV) and black queen cell virus (BQCV) were the only viruses detected, at frequencies of 38% and 66%, respectively. Varroa destructor was significantly more prevalent in colonies of European ancestry (p < 0.05), and the intensity of parasitism by V. destructor or infection by DWV and BQCV was also significantly higher in colonies of European descent than in African descent colonies (p < 0.01), although no genotype−parasite associations were found for N. ceranae. Additionally, significant and positive correlations were found between V. destructor and DWV levels, and the abundance of these pathogens was negatively correlated with the African ancestry of colonies (p < 0.01). However, there were no significant effects of environment on parasitism or infection intensity for the colonies of both genotypes. Therefore, it is concluded that the genotype of honey bee colonies, but not climate, influences their resistance to DWV, BQCV, and V. destructor.

Honey Bee (Apis mellifera) Immunity.

At the individual level, honey bees (Apis mellifera) rely on innate immunity, which operates through cellular and humoral mechanisms, to defend themselves against infectious agents and parasites. At the colony level, honey bees have developed collective defense mechanisms against pathogens and pests, such as hygienic and grooming behaviors. An understanding of the immune responses of honey bees is critical to implement strategies to reduce mortality and increase colony productivity. The major components and mechanisms of individual and social immunity of honey bees are discussed in this review.

The Combined Effects of Varroa destructor Parasitism and Exposure to Neonicotinoids Affects Honey Bee (Apis mellifera L.) Memory and Gene Expression.

Honey bees (Apis mellifera L.) are exposed biotic and abiotic stressors but little is known about their combined effect and impact on neural processes such as learning and memory, which could affect behaviours that are important for individual and colony survival. This study measured memory with the proboscis extension response (PER) assay as well as the expression of neural genes in bees chronically exposed to three different sublethal doses of the insecticide clothianidin and/or the parasitic mite Varroa destructor. The proportion of bees that positively responded to PER at 24 and 48 h post-training (hpt) was significantly reduced when exposed to clothianidin. V. destructor parasitism reduced the proportion of bees that responded to PER at 48 hpt. Combined effects between the lowest clothianidin dose and V. destructor for the proportion of bees that responded to PER were found at 24 hpt. Clothianidin, V. destructor and their combination differentially affected the expression of the neural-related genes, AmNrx-1 (neurexin), AmNlg-1 (neuroligin), and AmAChE-2 (acetylcholinesterase). Different doses of clothianidin down-regulated or up-regulated the genes, whereas V. destructor tended to have a down-regulatory effect. It appears that clothianidin and V. destructor affected neural processes in honey bees through different mechanisms.

Interaction of Varroa destructor and Sublethal Clothianidin Doses during the Larval Stage on Subsequent Adult Honey Bee (Apis mellifera L.) Health, Cellular Immunity, Deformed Wing Virus Levels and Differential Gene Expression.

Honeybees (Apis mellifera L.) are exposed to many parasites, but little is known about interactions with abiotic stressors on their health, particularly when affected as larvae. Larvae were exposed singly and in combination to the parasitic mite Varroa destructor and three sublethal doses of the neonicotinoid insecticide clothianidin to evaluate their effects on survivorship, weight, haemocyte counts, deformed wing virus (DWV) levels and gene expression of the adult bees that subsequently developed. Clothianidin significantly reduced bee weight at the highest dose and was associated with an increase in haemocyte counts at the lowest dose, whereas V. destructor parasitism increased DWV levels, reduced bee emergence, lowered weight and reduced haemocyte counts. An interaction between the two stressors was observed for weight at emergence. Among the differentially expressed genes (DEGs), V. destructor infestation resulted in broader down-regulatory effects related to immunity that was often shared with the combined stressors, while clothianidin resulted in a broader up-regulatory effect more related to central metabolic pathways that was often shared with the combined stressors. Parasites and abiotic stressors can have complex interactions, including additive effects on reduced weight, number of up-regulated DEGs and biological pathways associated with metabolism.

Interaction of field realistic doses of clothianidin and Varroa destructor parasitism on adult honey bee (Apis mellifera L.) health and neural gene expression, and antagonistic effects on differentially expressed genes.

While many studies have examined the effects of neonicotinoid insecticides and the parasitic mite Varroa destructor on honey bees (Apis mellifera), more information on the combined effects of such stressors on gene expression, including neural related genes, and their impact on biological pathways is needed. This study analyzed the effects of field realistic concentrations of the neonicotinoid clothianidin on adult bees infested and not infested with V. destructor over 21 consecutive days and then determined bee survivorship, weight, deformed wing virus (DWV) levels and gene expression. V. destructor parasitism with or without clothianidin exposure was significantly associated with decreased survivorship, weight loss and higher DWV levels, while clothianidin exposure was only associated with higher levels of DWV. Expression analysis of the neural genes AmNlg-1, BlCh and AmAChE-2 showed that V. destructor caused a significant down-regulation of all of them, whereas clothianidin caused a significant down-regulation of only AmNrx-1 and BlCh. An interaction was only detected for AmNrx-1 expression. RNAseq analysis showed that clothianidin exposure resulted in 6.5 times more up-regulated differentially expressed genes (DEGs) than V. destructor alone and 123 times more than clothianidin combined with V. destructor. Similar results were obtained with down-regulated DEGs, except for a higher number of DEGs shared between V. destructor and the combined stressors. KEGG (Kyoto Encyclopedia of Genes and Genomes) biological pathway analysis of the DEGs showed that the stressor linked to the highest number of KEGG pathways was clothianidin, followed by V. destructor, and then considerably fewer number of KEGG pathways with the combined stressors. The reduced numbers of DEGs and KEGG pathways associated with the DEGs for the combined stressors compared to the stressors alone indicates that the interaction of the stressors is not additive or synergistic, but antagonistic. The possible implications of the antagonistic effect on the number of DEGs are discussed.

Evaluation of Dry and Wet Formulations of Oxalic Acid, Thymol, and Oregano Oil for Varroa Mite (Acari: Varroidae) Control in Honey Bee (Hymenoptera: Apidae) Colonies.

The efficacy and safety of dry and wet formulations of three nonsynthetic compounds, oxalic acid (OA), thymol (T), and oregano oil (OO), for the control of Varroa destructor Anderson and Trueman infestations in honey bee (Apis mellifera Linnaeus) colonies were determined. The treatments were OA in dust, OA diluted in glycerin solvent embedded in a towel, T in dust, T in glycerin solvent and towel, OO in dry microcapsules, OO in glycerin solvent and towel, and the control. The treatments were applied weekly for 4 wk during the fall season. The rates of acaricide efficacy, weekly mite fall, bee mortality, colony survivorship, and strength, were determined for each of the treatments. All formulations, with the exemption of OO microcapsules, were effective at controlling infestations of V. destructor. The most effective formulations were T dust (96.6%), T glycerin (92.4%), and OA glycerin (79%). More than 85% of the mites were killed during the first 2 wk of treatment with T formulations, compared to less than 30% for the OA glycerin formulation. The lowest efficacy rate was for OO microcapsules (21.3%), and the only treatment that significantly increased bee mortality was OA glycerin. The rates of winter colony survival and honey bee populations were related to the varroacidal efficacy of the formulations. The implications of these findings are discussed.

Selective Breeding for Low and High Varroa destructor Growth in Honey Bee (Apis mellifera) Colonies: Initial Results of Two Generations.

After two years of bidirectional selection for low and high rates of Varroa destructor population growth (LVG and HVG, respectively) in honey bee (Apis mellifera) colonies in Ontario, Canada, significant differences between the two genotypes were observed. LVG colonies had V. destructor population increases over the summer of 1.7 fold compared to 9.6 fold for HVG colonies by Generation 2. Additionally, HVG colonies had significantly higher mite infestation rates in adult bees compared to LVG colonies for both selected generations. DWV prevalence and levels were significantly higher in HVG colonies than in LVG colonies in Generation 1 but not in Generation 2. Winter mortality rates of Generation 1 colonies were significantly different at 26% and 14% for the HVG and LVG genotypes, respectively. The results of this study thus far indicate that selection for LVG may result in colonies with lower V. destructor infestation rates, lower prevalence, and levels of DWV and higher colony winter survivorship. Future work will focus on determining what mechanisms are responsible for the genotypic differences, estimating genetic parameters, and molecular analyses of the genotypes to identify candidate genes associated with resistance to V. destructor and DWV that could potentially be used for marker-assisted selection.

Seasonality of Nosema ceranae Infections and Their Relationship with Honey Bee Populations, Food Stores, and Survivorship in a North American Region.

Nosema ceranae is an emerging pathogen of the western honey bee (Apis mellifera L.), and thus its seasonality and impact on bee colonies is not sufficiently documented for North America. This study was conducted to determine the infection intensity, prevalence, and viability of N. ceranae in >200 honey bee colonies during spring, summer, and fall, in a North American region. We also determined the relationship of N. ceranae infections with colony populations, food stores, bee survivorship, and overwinter colony mortality. The highest rates of N. ceranae infection, prevalence, and spore viability were found in the spring and summer, while the lowest were recorded in the fall. N. ceranae spore viability was significantly correlated with its prevalence and infection intensity in bees. Threshold to high levels of N. ceranae infections (>1,000,000 spores/bee) were significantly associated with reduced bee populations and food stores in colonies. Furthermore, worker bee survivorship was significantly reduced by N. ceranae infections, although no association between N. ceranae and winter colony mortality was found. It is concluded that N. ceranae infections are highest in spring and summer and may be detrimental to honey bee populations and colony productivity. Our results support the notion that treatment is justified when infections of N. ceranae exceed 1,000,000 spores/bee.

Nosema ceranae causes cellular immunosuppression and interacts with thiamethoxam to increase mortality in the stingless bee Melipona colimana.

The microsporidian parasite Nosema ceranae and neonicotinoid insecticides affect the health of honey bees (Apis mellifera). However, there is limited information about the effect of these stressors on other pollinators such as stingless bees (Hymenoptera: Meliponini). We examined the separate and combined effects of N. ceranae and the neonicotinoid thiamethoxam at field-exposure levels on the survivorship and cellular immunity (hemocyte concentration) of the stingless bee Melipona colimana. Newly-emerged bees were subjected to four treatments provided in sucrose syrup: N. ceranae spores, thiamethoxam, thiamethoxam and N. ceranae, and control (bees receiving only syrup). N. ceranae developed infections of > 467,000 spores/bee in the group treated with spores only. However, in the bees subjected to both stressors, infections were < 143,000 spores/bee, likely due to an inhibitory effect of thiamethoxam on the microsporidium. N. ceranae infections did not affect bee survivorship, but thiamethoxam plus N. ceranae significantly increased mortality. Hemocyte counts were significantly lower in N. ceranae infected-bees than in the other treatments. These results suggest that N. ceranae may infect, proliferate and cause cellular immunosuppression in stingless bees, that exposure to sublethal thiamethoxam concentrations is toxic to M. colimana when infected with N. ceranae, and that thiamethoxam restrains N. ceranae proliferation. These findings have implications on pollinators' conservation.

Effects of sublethal doses of clothianidin and/or V. destructor on honey bee (Apis mellifera) self-grooming behavior and associated gene expression.

Little is known about the combined effects of stressors on social immunity of honey bees (Apis mellifera) and related gene expression. The interaction between sublethal doses of a neurotoxin, clothianidin, and the ectoparasite, Varroa destructor, was examined by measuring differentially expressed genes (DEGs) in brains, deformed wing virus (DWV) and the proportion and intensity of self-grooming. Evidence for an interaction was observed between the stressors in a reduction in the proportion of intense groomers. Only the lowest dose of clothianidin alone reduced the proportion of self-groomers and increased DWV levels. V. destructor shared a higher proportion of DEGs with the combined stressors compared to clothianidin, indicating that the effects of V. destructor were more pervasive than those of clothianidin when they were combined. The number of up-regulated DEGs were reduced with the combined stressors compared to clothianidin alone, suggesting an interference with the impacts of clothianidin. Clothianidin and V. destructor affected DEGs from different biological pathways but shared impacts on pathways related to neurodegenerative disorders, like Alzheimer's, which could be related to neurological dysfunction and may explain their negative impacts on grooming. This study shows that the combination of clothianidin and V. destructor resulted in a complex and non-additive interaction.